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Acta Agron Sin ›› 2016, Vol. 42 ›› Issue (03): 389-398.doi: 10.3724/SP.J.1006.2016.00389

• CROP GENETICS & BREEDING · GERMPLASM RESOURCES · MOLECULAR GENETICS • Previous Articles     Next Articles

Cloning of 6-SFT Gene from Leymus racemosus and Analysis of Tolerance to Drought and Cold Stresses in Transgenic Tobacco

HE Xiao-Lan1,WANG Jian-Wei2,LI Wen-Xu3,CHEN Zhen-Zhen1,ZHAO Ji-Xin1,WU Jun1,WANG Zhong-Hua1,CHEN Xin-Hong1,*   

  1. 1 Shaanxi Provincial Key Laboratory of Plant Genetic Engineering Breeding / College of Agronomy, Northwest A&F University, Yangling 712100, China; 2 College of Environment and Life Science, Kaili University, Kaili 556011, China; 3 Wheat Research Institute, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China
  • Received:2015-07-22 Revised:2015-11-20 Online:2016-03-12 Published:2015-12-18
  • Contact: 陈新宏, E-mail: cxh2089@126.com; Tel: 029-87082854 E-mail:helingzhi123@126.com
  • Supported by:

    This study was supported by the National Natural Science Foundation of China (31571650) and the Tang Zhong-Ying Breeding Funding Project of the Northwest A&F University.

Abstract:

The fructan biosynthesis enzyme (6-SFT) plays an important role in plant responses to abiotic stresses. In this study, a full-length cDNA encoding sucrose:fructan-6-fructosyltransferase, designated as Lr-6-SFT (GenBank accession No. KT387273), was cloned from Leymus racemosus (2n = 4x = 28, NsNsXmXm) using reverse transcriptase PCR (RT-PCR) and rapid-amplification of cDNA ends (RACE) techniques. The full-length open reading frame comprises 1863 bp and encodes 620 amino acids. The predicted protein structure of the gene containes a conserved fructosyltransferase domain. Multiple sequence alignment and phylogenetic analysis showed that the Lr-6-SFT protein shared high similarity with 6-SFT proteins from Psathyrostachys huashanica, Triticum aestivum, Aegilops searsii, and Hordeum vulgare subsp. vulgare. The Lr-6-SFT gene was transferred into tobacco (Nicotiana tabacum L. cv. W38) via Agrobacterium-mediated transformation. The screened plants were tested by PCR and RT-PCR, and the transgenic tobacco plants exhibited much higher tolerance to drought and cold compared with the non-transgenic plants. Under drought and cold stresses, the Lr-6-SFT expressions were associated with the increased accumulation of stored carbohydrate and proline and the decreased malondialdehyde storage. These results suggest that Lr-6-SFT is a typical member of the glycoside hydrolase 32 (GH32) family and may be linked to enhanced tolerance to drought and cold stresses.

Key words: 6-SFT gene, Drought, Freezing, Leymus racemosus, Physiological indices, Transgenic tobacco

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